Figure 5. Dietary MSC counteracted the effects of NMU on acetylation of BMAL1 and H3K9 in Per2 promoter in mammary glands of NMU-treated rats in vivo.

ChIP assay was performed with anti-AcBMAL1 or anti-AcH3K9 antibody and specific primer set against Per2 E-box or Exon1 in Per2 promoter. On day 30 post-exposure, three rats in each group were sacrificed every 4 hours over 24 hour, beginning at 7 AM (ZT0). Pooled mammary tissues from three rats per time per group subjected to ChIP. A. Schematic representation of the Per2 promoter illustrates the regions of PCR amplicons against E-box binding motif and Exon1 in ChIP; B. AcBMAL1 levels; C. AcH3K9 levels. Upper, E-box; middle, Exon1; lower, input control.